Determination of vitamin B6 in foods by HPLC — a collaborative study

Bergaentzlé, Martine; Arella, F; Bourguignon, J.B.; Hasselmann, C.

doi:10.1016/0308-8146(94)p4185-i

Cited by 33 publications

(18 citation statements)

References 7 publications

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“…Os procedimentos bioanalíticos e microbiológicos são trabalhosos e consomem longos tempos de análise 3,4 , enquanto que os métodos químicos apresentam maior precisão e menor tempo de execução 5,6 . Os procedimentos químicos mais empregados incluem a cromatografia gasosa 7 , cromatografia líquida de alta eficiên-cia (CLAE) [8][9][10][11] , espectrofotometria no UV-visível 12-14 , potenciometria 15,16 , espectrofluorimetria 17,18 , espectrofotometria em sistema por injeção em fluxo 19,20 procedimentos em sistemas por injeção em fluxo para determinação de vitamina B 1 em formulações farmacêuticas com detecção espectrofotométrica 24 e turbidimétrica 25 . Em continuidade a esses estudos, um procedimento em fluxo com detecção espectrofométrica é proposto para a determinação direta de vitamina B 6 (cloridrato de piridoxina) em formulações farmacêuticas, sem a necessidade prévia de separar essa vitamina por extração por solventes ou outro procedimento de separação.…”

Section: Introductionunclassified

Determinação espectrofotométrica por injeção em fluxo de vitamina B6 (piridoxina) em formulações farmacêuticas

Aniceto¹,

Fatibello‐Filho²

1999

Quím. Nova

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Keywords: vitamin B 6 ; flow injection spectrophotometry; pharmaceutical preparations. ARTIGO INTRODUÇÃOVitamina B 6 (cloridrato de piridoxina) é um pó cristalino branco, inodoro e relativamente estável ao ar e à luz. Suas soluções aquosas são mais estáveis em valores de pH menores que 5, tornando-se menos estáveis em valores de pH maiores que 7, especialmente quando irradiadas com luz na região do UV-Visível 1 . Vitamina B 6 está amplamente distribuída na natureza, sendo as fontes de sua obtenção o fígado, farelo de cereais, levedura, melaço bruto de cana e germe de trigo. Essa vitamina consiste de uma mistura de piridoxina, piridoxal e piridoxamina (Figura 1), que são normalmente interconvertidas no organismo. Desses compostos, a piridoxina é a mais estável sendo normalmente usada nas preparações farmacêuticas na forma de sais como cloridrato (Adermina), ascorbato, aspartato e piridofilina. São também utilizados vários derivados análogos da piridoxina, tais como: (a) iodometilato de piridoxina, usado por via oral ou parenteral; (b) piritinol (Encefabol), desprovido de atividade neurotrófica; (c) 3-lauroil-4,5-dialcetilpiridoxina, indicada para o tratamento de dermatoses e (d) piridoxilato, um precursor da piridoxina. No organismo a piridoxina converte-se em fosfato de piridoxal, que atua como coenzima de cerca de 60 enzimas, a maioria relacionada com o metabolismo de proteí-nas e aminoácidos. Essa vitamina desempenha importante papel na síntese de neurotransmissores como a noradrenalina, dopamina, serotonina e histamina. Participa também de reações de degradação de aminoácidos, em que um dos produtos finais é a acetil-coenzima A, necessária à produção de energia e à síntese de proteínas, lipídios e acetilcolina 1,2 . O fosfato de piridoxal atua como coenzima na primeira etapa da síntese de esfingosina, substância que ocupa posição chave no metabolismo dos esfingolipídios, componentes essenciais nas membranas celulares das bainhas de mielina. Uma vez que os esfingolipídios têm renovação metabólica muito rápida, a preservação da integridade estrutural e funcional do sistema nervoso requer a síntese constante de esfingosina, dependente de vitamina B 6 1,2 . A carência de piridoxina determina alterações na pele e mucosas, lesões seborréicas da face, glossite, estomatite; no sistema nervoso central e periférico, convulsões, depressão, neuropatia; na hematopoese, anemia microcítica hopocrômica, com reserva normal ou aumentada de ferro (anemia sideroblástica) 1,2 . Existem vários procedimentos para a determinação de vitamina B 6 descritos na literatura 3-23 , destacando-se os bioanalíticos, microbiológicos e químicos. Os procedimentos bioanalíticos e microbiológicos são trabalhosos e consomem longos tempos de análise 3,4 , enquanto que os métodos químicos apresentam maior precisão e menor tempo de execução 5,6 . Os procedimentos químicos mais empregados incluem a cromatografia gasosa 7 , cromatografia líquida de alta eficiên-cia (CLAE)

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Section: Introductionunclassified

Determinação espectrofotométrica por injeção em fluxo de vitamina B6 (piridoxina) em formulações farmacêuticas

Aniceto¹,

Fatibello‐Filho²

1999

Quím. Nova

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“…The advantage of this excellent pre-column derivatization was that only one vitamer, pyridoxine, needed to be finely measured. The validity of the method was further tested by a collaborative study in which good repeatability and reproducibility was obtained (Bergaentzle et al 1995). This method has been put forward as an official method in France for vitamin B 6 determination in foods.…”

Section: Detectionmentioning

confidence: 99%

“…Thus prolonged incubation time, 12h to overnight at 37°C, to ensure the proper hydrolysis was recommended. An acid phosphatase treatment combined with conversion of pyridoxal and pyridoxamine into pyridoxine is well documented including the collaborative study parameters (Reitzer-Bergaentzle et al 1993, Bergaentzle et al 1995. Hydrolysis based on alkaline phosphatase has been carried out after perchloric acid extraction.…”

Section: Dephosphorylationmentioning

confidence: 99%

“…Acidic acetate solution was easily combined with the followed enzymatic hydrolysis procedure and the method was suitable for the simultaneous measurement of free B 1 , B 2 and B 6 vitamers in plasma and serum samples. The validity of the acetate extraction used by Reitzer-Bergaentzle et al (1993) preceeding the pre-column derivatization of B 6 vitamers into pyridoxine was further evaluated by the collaborative study performed in France (Bergaentzle et al 1995). A lower pH acetate buffer was used for multivitamin preparate; sample was sonicated with a dilute solution of acetic acid (0.5M) and triethylamine (40mM) at the pH of 3.6 (Lam et al 1984) or with liquid chromatography's mobile phase (acetonitrile -10mM phosphate buffer -triethylamine, 8:91.5:0.5, pH 3.8) (Maeda et al 1989).…”

Section: Metaphosphoric Acid (Mpa)mentioning

confidence: 99%

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HPLC analysis of vitamin B6 in foods

Ollilainen¹

1999

AFSci

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The objective of this work was to evaluate the methods for determination of vitamin B6 in foods. To achieve this, the literature review focused on sample treatment and liquid chromatographic analysis of vitamin B6 related compounds. In the experimental part, the chosen sample pretreatment and the high-performance liquid chromatographic (HPLC) method were validated, and used to produce vitamin B6 data on various food items commonly consumed in Finland. The main emphasis of the sample treatment was on the extraction efficiency and the maintenance of the original concentration profile of the vitamers. Several acid extraction procedures were tested for this purpose. Perchloric acid was chosen as the extraction agent. Routine food analysis was then performed using dilute ice-cold perchloric acid extraction followed by an internally standardized ion-paired reversed-phase liquid chromatography. Food samples were hydrolyzed with takadiastase and alkaline phosphatase enzymes, phosphorylated and glycosylated vitamers were quantitated before and after the enzymatic digestion. This procedure enabled the extraction of vitamin B6 compounds in their intact forms, and the measurement of free, phosphorylated and glycosylated forms. The maintenance of the concentration profile of the vitamers was verified by using 14C -labeled pyridoxal 5'-phosphate in the examination of the extraction procedure. The extraction efficiency and laboratory performance were confirmed by interlaboratory studies. Up-to-date data on vitamin B6 content of about fifty common food items was produced. The data includes the results from meat and poultry, fish and fish product, dairy product, cereal and vegetable, and ready-to-eat food samples. Free and phosphorylated vitamin B6 compounds were measured in all food groups, and the glycosylated vitamer fraction was analyzed in all plant-derived foods. The results obtained in this work showed that vitamin B6 content of nearly all foods of plant origin was mainly comprised of glycosidically bound pyridoxine derivatives. These bound analytes are normally not taken into account in traditional analytical methods, and food composition tables lack the data of glycosylated pyridoxine. The role of the glycosylated pyridoxines need to be clarified in terms of their analytical and physiological nature. If, as it is currently assumed, the availability of the bound forms is limited for humans, the role of vegetables, cereals and other foods of plant origin as a source of vitamin B6, as well as the analytical methods should be reassessed.;

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“…In addition, it is very probable that the inhibition of acid phosphatase by APS raise a complicated problem when evaluating phosphorylated compounds in foods containing APS. For example, the phosphorylated forms of vitamins B6 in foods are often estimated by hydrolyzing them with potato acid phosphatase 6,7) or alkaline phosphatase 8) and then measuring the free forms of the vitamin. In a preliminary experiment, we found that the phosphorylated forms of vitamin B6 in plant foods such as orange juice, lemon juice, broccoli, and banana (apple contains little of the phosphorylated forms), which are known to contain pectin, were incompletely (only 30¿60z) dephosphorylated by potato acid phosphatase, whereas they were completely hydrolyzed by the same amount of alkaline phosphatase, indicating that the phosphorylated forms of vitamin B6 were underestimated when potato acid phosphatase was used.…”

mentioning

confidence: 99%